Protein Seeding Detection
Protein Seeding Detection - Related Videos
04:11
A FRET Flow Cytometry Technique to Detect Tau-Seed Induced Reporter Protein Aggregation
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2025
The video demonstrates a fluorescence resonance energy transfer (FRET) flow cytometry assay to detect the seeding activity of protein aggregates isolated from biological samples. Mammalian cells expressing tau reporter proteins are incubated with liposome transduction complexes containing tau seeds. These seeds mediate the aggregation of the reporter proteins, leading to generating a FRET positive signal in the flow cytometer.
12:31
Sensitive Detection of Proteopathic Seeding Activity with FRET Flow Cytometry
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Cited by 95 •
2015
Cell-to-cell transfer of protein aggregates, or proteopathic seeds, may underlie the progression of pathology in neurodegenerative diseases. Here, a novel FRET flow cytometry assay is described that enables specific and sensitive detection of seeding activity from recombinant or biological samples.
09:00
Detection of Protein Ubiquitination
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Cited by 97 •
2009
Ubiquitination is a key posttranslational modification carried out by a set of three enzymes. Mutations of genes involved in this modification are associated with many different human diseases. Here, we describe protocols to detect protein ubiquitination in cultured cells in vivo and test tubes in vitro.
03:41
Fluorescence Anisotropy-Based Detection of Protein-Protein Interactions
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2025
In this video, we describe the fluorescence anisotropy technique to study the interactions between the fluorophore-tagged Shwachman-diamond syndrome (SBDS) protein and the elongation factor-like 1 GTPase (EFL1). On incubating SBDS proteins with gradually increasing concentrations of EFL1, a steady increase in anisotropy is observed, indicating a successful interaction between the two proteins.
07:42
In-vivo Detection of Protein-protein Interactions on Micro-patterned Surfaces
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Cited by 8 •
2010
This video shows experiments with subsequent analysis of protein-protein interactions by the use of micro-patterned surfaces. The approach offers the possibility to detect protein interactions in living cells and combines high throughput capabilities with the possibility to extract quantitative information.
FAQs
The recombinant protein substrate supplies normally folded protein that can be converted when a misfolded seed is present. This creates a controlled reaction system in which the sample’s seeding activity can be amplified rather than measured only at its original concentration. The substrate therefore connects the presence of an abnormal assembly to a detectable aggregation-associated fluorescence signal.
Repeated shaking promotes the seed-dependent aggregation process by repeatedly disturbing the reaction mixture. As aggregation proceeds, the assay generates fluorescence over time, allowing the reaction to be monitored dynamically rather than as a single endpoint. This time-dependent behavior helps distinguish reactions in which a sample initiates conversion from those lacking detectable seeding activity.
High analytical sensitivity allows these methods to identify very small amounts of misfolded protein assemblies in biological specimens. That capability is important because abnormal proteins may be present at low abundance while still providing information about disease-associated processes. Sensitive detection supports studies of pathogenic assemblies, their possible transmission, and the development of biomarker-based approaches.
A typical workflow combines a biological specimen with a recombinant protein substrate, then subjects the mixture to repeated shaking while monitoring fluorescence over time. The resulting signal reflects seed-dependent aggregation within the reaction. This workflow converts a difficult-to-measure abnormal protein signal in the original specimen into an amplified, measurable readout for research analysis.
Protein Seeding Detection supports investigations of prion-like agents and other pathogenic protein assemblies in biological specimens. Researchers can apply the methods to questions involving neurodegeneration, infection, transmission, and diagnostic biomarker development. Because the assay amplifies seeding activity, it also helps examine how abnormal proteins may spread and contribute to disease-associated processes.
Within immunology and infection research, these assays provide a way to investigate abnormal protein agents in biological specimens and to examine their relevance to infection-related transmission. The resulting measurements can support research on prion-like agents, pathogenic protein assemblies, and biomarker development, extending infection studies beyond conventional analysis of intact biological organisms or cells.